Analysis of LMP1 and TRAF1 domains important for M1-pUb attachment.A) 293 cells were co-transfected with FLAG-TRAF1, and with either wildtype LMP1 (WT), an LMP1 mutant that signal only from the TES2 domain (TES2), an LMP1 mutant that signals only from the TES1 domain (TES1), or an LMP1 double mutant (DM) that does not signal from either TES1 or TES2. Immuno-purified FLAG-TRAF1 complexes and whole cell lysates were blotted, as indicated. B) 293 cells were transfected with FLAG-tagged WT LMP1, LMP1 1–231, or LMP1 DM and untagged TRAF1. Purified FLAG complexes or lysates were blotted as indicated. C) 293 cells were co-transfected with untagged LMP1 and FLAG-tagged GFP, TRAF1 1–416, TRAF1 183–416, or TRAF1 264–416. FLAG complexes or lysates were blotted, as indicated. A-C are representative of triplicate experiments.
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